Fluorimetric HDAC Activity Assay Kit (Green)

Our Fluorimetric HDAC Activity Assay Kit provides a quick, convenient, and sensitive method for the detection of HDAC activity. This kit uses our non-peptide HDAC Green substrate that is much more sensitive than the peptide-based HDAC substrates such as Ac-RGK(Ac)-R110, Ac-RGK(Ac)-AMC and Ac-RGK(Ac)-AFC. In addition, HDAC Green substrate is also much more resistant to protease hydrolysis than other commercial peptide-based HDAC substrates. Our kit can be used for measuring HDAC activity in cell lysates or HDAC inhibitor screening with cell extracts or purified enzymes. The long wavelength emission of the HDAC Green substrate makes the assay less interfered from compounds and cell components. HDAC activity is monitored with excitation at 490 nm and emission at 525 nm.

Description:

Histone deacetylases (HDAC) are a class of enzymes that remove acetyl groups from a ε-N-acetyl lysine amino acid on a histone. Deacetylation restores the positive electric charge of the lysine amino acids, which increases the histone’s affinity to the negatively charged phosphate backbone of DNA. This process generally down-regulates DNA transcription by blocking the access of transcription factors. HDAC inhibitors are being studied as a treatment for cancer.

Broad Application: Can be used for quantifying HDAC in solutions and in cell extracts.Continuous: Easily adapted to automation without a separation step.Convenient: Formulated to have minimal hands-on time. No wash is required.Non-Radioactive: No special requirements for waste treatment.

The fluorescence in blank wells (with the assay buffer only) is used as the background fluorescence, and is subtracted from the values for those wells with the HDAC Green reactions. All fluorescence readings are expressed in relative fluorescence units (RFU).